Background: Prescription drug misuse and abuse is an established public health challenge, and young adults are particularly affected. There is a striking lack of real-time, naturalistic data collection assessing intentions to misuse and other precipitating factors at the time of actual misuse, leaving unknown under what conditions individuals are most likely to misuse prescription medications. Ecological momentary assessment (EMA) applications and protocols designed to capture this information would accelerate and expand the knowledge base and could directly contribute to prevention and treatment efforts.Objective: The objectives in the current study were to (1) describe the development and administration of a mobile application and EMA protocol designed to collect real-time factors associated with college students’ prescription drug misuse intentions and behaviors in daily life; (2) present completion rates, compliance, acceptability, and reactivity associated with the EMA protocol for participants who endorsed recent prescription drug misuse at screening (n = 300; i.e., risk group) and those who did not (n = 55; i.e., non-risk group); and (3) establish initial construct validity by linking the reports of misuse behavior in daily life collected via the EMA app to prescription drug misuse reported on a standard survey.Methods: An EMA data collection app and protocol were designed specifically to capture hypothesized contextual factors along with prescription drug misuse intentions and behaviors in daily life. Using the protocol, young-adult college students (N = 352) completed signal- and event-contingent reports over a 28-day period. When intention to misuse a prescription drug was endorsed, a brief follow-up prompt was sent 15 minutes later to collect participants’ indications of whether or not misuse had occurred.Results: Risk-group participants were significantly more likely than non-risk counterparts to endorse any prescription drug misuse intentions in daily life, to complete one or more follow-up report, and to endorse any prescription drug misuse behavior in daily life on the follow-ups (P-values < .001). Overall, participants demonstrated consistent engagement with the EMA procedures and returned an average of 74.5 reports (SD = 23.82, range 10-122). Participants in risk and non-risk groups did not differ in the number of reports they completed, the number of their reporting days, or their average completion rates (P-values > .10). Results indicated some evidence of reactivity to the momentary reporting procedure. Participants reported uniformly positive experiences and remained highly engaged throughout the reporting protocol and broader study.Conclusions: The novel EMA app and protocol provide an effective way to assess real-time factors associated with prescription drug misuse intentions and behavior in daily life. The resulting investigations offer the potential to provide highly translatable information for research and prevention efforts.